A kind of spider mitochondrial coiii gene complete sequence amplification primer and identification method

A COIII-F, amplification primer technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Low equipment requirements, shortened identification cycle, and the effect of overcoming difficulties

Active Publication Date: 2017-09-26
CHINA JILIANG UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, the current mitochondrial COIII Universal amplification primers for genes have not been reported
The lack of universal primers with high amplification efficiency severely hampers COIII Application of genes as molecular markers, thus engineered to specifically amplify spider mitochondria COIII The universal primer of the gene is of great significance to the rapid identification of spider species and the investigation and analysis of species diversity by molecular methods, and it also lays the foundation for the determination of the whole genome sequence and phylogenetic analysis of spider mitochondria

Method used

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  • A kind of spider mitochondrial coiii gene complete sequence amplification primer and identification method

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Embodiment 1

[0021] (1) Collection, identification and preservation of spider samples

[0022] The species of spiders used in the examples were all collected from the farmlands of Yuyao City or Tongxiang County, Zhejiang Province by trap method. The collected specimens were brought back to the laboratory for information registration, and all were identified with a stereo microscope to determine the species. The spider specimens were soaked in 100% alcohol and stored in a 4°C refrigerator for later use. The species of spiders collected include: Pseudomonas tarantula, Triflower spider, Golden spider, Cone-bellied octopus, Ornate octopus, Brown new orb spider, Maiwa spider, black fly tiger, big-bellied garden spider, cat jumping spider.

[0023] Extraction of Total DNA from Spider Genome

[0024] DNA extraction kits were used to extract the total genomic DNA of different species of spiders. To avoid contamination, the head and abdomen were discarded, and DNA was extracted only from the leg...

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Abstract

The invention discloses a primer for amplifying the full sequence of the mitochondrial COIII gene of a spider, and a method for quickly identifying spider species by using the sequencing information of the full sequence, including the extraction of the total DNA of the spider genome, the design of the primer for amplifying the full sequence of the COIII gene, and PCR Amplification, sequencing and rapid identification of spider species based on the full sequence of the COIII gene. The universal amplification primer of the present invention can rapidly and efficiently amplify the complete sequence of the mitochondrial COIII gene of various spiders, and provide a powerful tool for rapid identification of spider species, analysis of population diversity, protection and utilization of germplasm resources, etc. , It also lays the foundation for the analysis and research of spider mitochondrial genome sequencing and the evolution of different taxonomic levels.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer for amplifying the complete sequence of a spider mitochondrial cytochrome C oxidase 3 (cytochrome oxidase subunits 3; COIII) gene and a method for identifying spider species. Background technique [0002] Spiders are an important predatory natural enemy of pests and play an important role and status in the process of integrated pest management. Pay attention to the protection of the seed bank and promote the reconstruction of the natural enemy community, which can effectively control the occurrence and damage of many agricultural pests, such as planthoppers, leafhoppers, stem borers, and cotton aphids. Rapid and accurate species identification is the necessary premise and basis for the research on the protection and utilization of spiders. However, the traditional classification and identification methods based on appearance and anatomical features are not only ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 王正亮俞晓平李超
Owner CHINA JILIANG UNIV
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